恒河猴肾上腺皮质的显微与超微结构

应用光镜和透射电镜观察，恒河猴肾上腺皮质可分为球状带、束状带和网状带。其特点是：（1）被膜较发达，有时可伸入球状带将其细胞分隔为团块。（2）球状带有时呈指状指入束状带和网状带，其内可见嗜铬细胞；球状带细胞可分为明、暗2种类型，他们排列成团球状、滤泡状、索状和不规则形。（3）网状带较发达，有的可分为内、外2个区；有时网状带呈刺状伸入束状带；有时网状带细胞可见于被膜下或髓质内。（4）有时部分被膜连同皮质一起呈指状伸入髓质内。（5）皮质细胞均含丰富的溶酶体、线粒体和内质网，但不同部位的细胞所含的脂滴在数量、大小、形态和电子密度等方面存在差异。     

Morphokinese der Nebennierenrinde beim Rind nach ein-oder mehrfacher Glukokortikoidapplikation zur Geburtseinleitung

Morphometrisch untersucht wurden die Nebennieren von 27 Kühen mit und ohne medikamentelle Geburtseinleitung bzw. nicht tragender Kontrolltiere (Gruppe I (n = 7): 10 mg Triamcirwlon −6,5 d ante partum und 10 mg Flumethason −30 h ante partum; Gruppe II (n = 6): 1 ml NaCl (Placebo) −6,5 d ante partum und 10 mg Flumethason −30h ante partum; Gruppe III (n = 4): tragend, keine Vorbehandlung; Gruppe IV (n = 10): nicht tragende Kontrolltiere). Eine Geburtsinduktion mit Glukokortikoiden führt zu einer signifikanten Verkleinerung der Kerngröβen sowie einer Zunahme der Zellkernzahl pro definierter Flächeneinheit in der Zona fasciculata und einer signifikanten Verschmälerung von Zona fasciculate/Zona reticularis. Diese Befunde werden als medikamentell induzierte regressive adrenokortikale Transformation interpretiert.
Contents: Morphokinesis of the adrenal cortex in cattle after induced parturition by single or repeated glucocorticoid application
Adrenal glands of 27 cows with or without induced parturition and non-pregnant controls were investigated morphometrically (Group I (n = 7): 10 mg Triamcinolone - 6,5 d ante partum and 10 mg Flumethasone −30 h ante partum; group II (n = 6): 1 ml NaCl (placebo) - 6,5 d ante partum and 10 mg Flumethasone −30 h ante partum; group III (n = 4): pregnant, nopretreatment; group N(n= 10): non-pregnant controls). Pretreatment with glucocorticoids results in a significant reduction of nuclear sizes and an increase in number of nuclei per area unit defined in the zona fasciculata, combined with a significant decrease of the width of zona fasciculate/zona reticularis. These findings are interpreted as indicator for a medicamentally induced regressive adrenocortical transformation.     

Adrenal Cortex of the Dog (Canis familiaris): I. Histomorphologic Changes during Growth,Maturity, and Aging1

The adrenal glands of 56 dogs ranging in age from birth to 13.6 years were examined for a determination of normal zonal and cellular morphology patterns throughout development, maturity and aging. Routine light microscopic techniques as well as lipid and cholesterol staining were used. The zona arcuata is described as developing between the fibrous and cellular layers of the capsule forming the cortical blastema layer, a zona biproliferativa. It is proposed that both centrifugal and centripetal migration of cortical parenchyma can result. The formation of accessory nodules is described as compensatory hyperplasia. Histomorphometric analysis revealed growth in the zona fasciculata to be by hyperplasia and the zonal volume percentages to be arcuata, 27%; fasciculata, 50%; reticularis, 23%.     

Histological evaluation of the adrenal glands of seven dogs with hyperadrenocorticism treated with trilostane

The lesions in the adrenal glands of seven dogs with hyperadrenocorticism that had been treated with trilostane were studied histologically. The glands of the six dogs with pituitary-dependent hyperadrenocorticism had moderate to severe cortical hyperplasia that was either diffuse or nodular. The lesions were more pronounced in the zona fasciculata than in the zona reticularis, and the zona glomerulosa was normal. In the dog with a functional adrenal tumour the non-tumour bearing adrenal gland showed mild nodular hyperplasia. Five of the seven dogs had variable degrees of adrenal necrosis, which was severe in two of them. The terminal deoxynucleotidyl transferase-mediated DUTP nick-end labelling (TUNEL) reaction specified areas of cell death as apoptosis in three of the dogs, and was positive in one of the dogs without visible areas of cell death. There were variable degrees of cortical haemorrhage in three of the dogs. In some of the dogs the lesions were severe enough to lead to hypoadrenocorticism.     

Adrenal cortical necrosis caused by T-2 mycotoxicosis in female, but not male, mice

Experimentally, adrenal cortical parenchymal cell necrosis was induced by T-2 mycotoxin in female, but not male, mice. The lesion occurred in the adrenal glands in 11 of 11 female and 0 of 10 male mice given a nose-only aerosol exposure to T-2 mycotoxin. The necrosis, restricted to the zona fasciculata, began at the X zone interface and extended peripherally to involve 15% to 70% of the zone. Both light and transmission electron microscopies were used to determine whether the cellular and subcellular damage involved parenchymal cells of the zona fasciculata. Extensive necrosis of cortical thymocytes and necrosis of lymphoid cells in follicles of the spleen, lymph nodes, and intestine-associated lymphoid tissue were observed in both sexes. This is the first report to describe adrenal gland necrosis associated with exposure to T-2 mycotoxin.     

Suspected spontaneous hyperadrenocorticism in a young experimental beagle dog

A 6-month-old female beagle dog, assigned to the low-dose group in a toxicity study, was evaluated for compound toxicity, and spontaneous hyperadrenocorticism was suspected. The animal had an externally apparent distended abdomen on clinical examination upon arrival. Pre-dose clinical pathology showed slightly higher erythroid parameters and stress leukogram on hematology; plasma biochemistry showed higher total protein, gamma-glutamyl transferase, total cholesterol, and triglyceride levels than the reference data. On necropsy, a prominent increase in adipose tissues of the subcutis and abdomen and increased weight of the adrenal gland and liver were observed. Histopathology revealed diffuse hyperplasia of adrenocortical cells in the zona fasciculata and reticularis, cortical atrophy of the thymus, and abundant glycogen accumulation in the hepatocytes. These findings were incidental and not test-substance-related. Electron microscopy of the adrenocortical cells in the zona fasciculata revealed decreased typical translucent lipid droplets, increased electron-dense lipid droplets, and abundant smooth endoplasmic reticulum and lysosomes. Additionally, increased numbers of various sizes and forms of mitochondria with tubular, vesicular, or lamellar cristae compared to that of normal animals were observed. These ultrastructural characteristics of the adrenocortical cells suggested hyperfunction. The pre-dose plasma cortisol levels were slightly higher than those of other females assigned to the toxicity study, while plasma adrenocorticotropic hormone levels were within the normal range. These findings indicate that hyperadrenocorticism is a possible cause of the systemic changes in this case.     

Differences in adrenal morphology in male Dark Agouti, Sprague-Dawley and Wistar rats.

There is little published information describing the differences in adrenal structure between strains of a single species despite quite well known functional differences. In this paper we report morphological differences in the adrenal glands between three strains of laboratory rat; Dark Agouti (DA), Sprague-Dawley (SD) and Wistar (W). Significant differences in adrenal weights (as a percentage of body weight) were not detected between any of the groups. While there were no significant differences in the volume of medullary or zonae glomerulosa or fasciculata tissue, the volume fraction for the zona reticularis was significantly smaller in DAs compared to the other strains. The functional significance of these differences is unknown. However, it is suggested that the reduced volume of zona reticularis tissue may contribute to a reduced capacity for glucocorticoid synthesis or storage.     

Attempts to influence birth in swine with dexamethasone. 2. Effect of DXMS on adrenal mass and cell nuclear volume of the fascicular zone in piglets

The adrenal weights of spontaneously born piglets, immediately after parturition, were significantly higher than those of piglets from sows in whom parturition had been induced by DXMS. The differences between mean values went down with DXMS treatment started later in advanced pregnancy. Mean absolute and relative adrenal weights of the DXMS groups came closer to those of the untreated controls on the tenth day of piglet age. Cell nucleus volumes of the zona fasciculata were normal in all groups and did not reveal any influence of DXMS treatment in any of the newborn piglets. The mean values were 89.9 micron3 in the control group and 87.6 micron3 in experimental group V (75 mg DXMS per animal and die on 109th through 111th days of pregnancy).     

Tissue gamma-glutamyl transpeptidase activity and hepatic ultrastructural alterations in dogs with experimentally induced glucocorticoid hepatopathy

Glucocorticoid hepatopathy was induced in 2 Beagle dogs with IM injections of prednisone, 4.4 mg/kg, given once daily for 14 consecutive days. Serum enzymatic activities were monitored and percutaneous hepatic biopsy materials were collected. A placebo-treated group of 2 Beagle dogs was given saline solution injections. Treated dogs showed clinical signs of Cushing's disease and alterations in mean serum enzymatic activities consistent with glucocorticoid hepatopathy. The gamma-glutamyl transpeptidase activities in placebo-treated dogs were highest for kidney, less (but marked) for pancreas, and least for liver, gallbladder, jejunum, duodenum, spleen, heart, lung, skeletal muscle, and erythrocytes. In treated dogs, the gamma-glutamyl transpeptidase activities were highest for pancreas and were decreased for kidney and slightly increased for the other tissues. Results of hepatic biopsy and necropsy examinations of treated dogs showed swollen, pale, friable hepatic parenchyma with hepatocytic cytoplasmic vacuolation and glycogen accumulation. Extrahepatic lesions included gastric ulceration, catarrhal enteritis, splenic and lymph node atrophy and lymphocytolysis, and vacuolation of adrenal cortical cells in the zona fasciculata. Ultrastructural study of liver from dogs with glucocorticoid hepatopathy indicated that the principle alterations were abundant glycogen accumulation and few mitochondria in hepatocytes.     

Acaricide treatment prevents adrenocortical hyperplasia as a long-term stress reaction to psoroptic mange in cattle

In cattle, infestation with Psoroptes ovis mites may cause severe dermatitis (psoroptic mange) which compromises the health and welfare of the animals and may lead to significant economic losses. To investigate yet undocumented effects of psoroptic mange mite infestations and how successful therapy promotes animal health, the present study examined alterations of the skin, lymph nodes and adrenal glands of P. ovis infested Fleckvieh (Simmental) bulls treated with either ivermectin long-acting injection (IVM LAI; IVOMEC^® GOLD, Merial; 3.15% ivermectin w/v) or saline (n = 16 each). Approximately 8 weeks subsequent to experimental infestation with P. ovis, the bulls had developed mange and were administered either IVM LAI or saline once at 1 mL/50 kg body weight by subcutaneous injection. Mite counts were conducted in weekly intervals for determination of efficacy of treatment, and following humane euthanasia of the animals 8 weeks after treatment, skin samples from affected (mangy or previously mangy) and unaffected areas, prescapular lymph nodes and adrenal glands were collected for gross and pathohistological examination. In addition, four age-matching, uninfested Simmental bulls were sampled as controls for comparison.No P. ovis mites were detected on any IVM LAI-treated bull after 28 days following treatment whereas saline-treated bulls maintained infestation throughout the study. At sampling (approximately 16 weeks after experimental infestation and 8 weeks following saline or IVM LAI treatment), saline-treated bulls displayed a severe, exsudative dermatitis with significantly increased skin thickness and inflammatory cell infiltration, significantly enlarged, hyperplastic prescapular lymph nodes, as well as significantly increased adrenal gland weights and volumes as compared to P. ovis-infested, IVM LAI-treated bulls and uninfested controls. Quantitative stereological analysis revealed that the adrenal gland enlargement in P. ovis-infested, saline-treated bulls was due to a selective increase of the volume of the zona fasciculata in the adrenal cortex. Compared to uninfested controls and P. ovis-infested, IVM LAI-treated bulls, the number of epithelial cells in the zona fasciculata was significantly increased in P. ovis-infested, saline-treated bulls, while the zona fasciculata cell volumes did not differ between the three groups of cattle. While the single point determination of serum cortisol concentrations did not reveal significant differences between the three groups of cattle at tissue sampling, the hyperplastic growth of the adrenal cortex in the P. ovis-infested, saline-treated bulls provides morphologic evidence that a chronic stress reaction is one consequence of mange mite infestations that can be prevented by efficacious acaricidal treatment.     

Histocytological specificities of adrenal cortex in the New World Monkeys, Aotus lemurinus and Saimiri boliviensis

The New World monkey Aotus spp. (night monkeys) are expected for use of valuable experimental animal with the close species of Saimiri spp. (squirrel monkeys). Saimiri is known to show spontaneous hypercortisolemia, although few reports in Aotus. We compared basic states of blood steroid hormones and histological structure of the adrenal glands in two monkeys. Serum cortisol and ACTH levels were statistically lower in Aotus than Saimiri. Conversely, Aotus adrenocortical area showed significant enlargement, especially at the zona fasciculata. Electron microscopic observation at Aotus fasciculata cells revealed notable accumulation of large lipid droplets and irregular shapes of the mitochondrial cristae. These results suggest potential differences in cellular activities for steroidogenesis between Aotus and Saimiri and experimental usefulness in adrenocortical physiology and pathological models.     

Expression of receptors for luteinizing hormone,gastric-inhibitory polypeptide,and vasopressin in normal adrenal glands and cortisol-secreting adrenocortical tumors in dogs

Hypercortisolism caused by an adrenocortical tumor (AT) results from adrenocorticotropic hormone (ACTH)-independent hypersecretion of glucocorticoids. Studies in humans demonstrate that steroidogenesis in ATs may be stimulated by ectopic or overexpressed eutopic G protein-coupled receptors. We report on a screening of 23 surgically removed, cortisol-secreting ATs for the expression of receptors for luteinizing hormone (LH), gastric-inhibitory polypeptide (GIP), and vasopressin (V_1a, V_1b, and V₂). Normal adrenal glands served as control tissues. Abundance of mRNA for these receptors was quantified using quantitative polymerase chain reaction (QPCR), and the presence and localization of these receptors were determined by immunohistochemistry. In both normal adrenal glands and ATs, mRNA encoding for all receptors was present, although the expression abundance of the V_1b receptor was very low. The mRNA expression abundance for GIP and V₂ receptors in ATs were significantly lower (0.03 and 0.01, respectively) than in normal adrenal glands. The zona fasciculata of normal adrenal glands stained immunonegative for the GIP receptor. In contrast, islands of GIP receptor-immunopositive cells were detected in about half of the ATs. The zona fasciculata of both normal adrenal glands and AT tissue were immunopositive for LH receptor; in ATs in a homogenous or heterogenous pattern. In normal adrenal glands, no immunolabeling for V_1bR and V₂ receptor was present, but in ATs, V₂ receptor-immunopositive cells were detected. In conclusion, QPCR analysis did not reveal overexpression of LH, GIP, V_1a, V_1b, or V₂ receptors in the ATs. However, the ectopic expression of GIP and V₂ receptor proteins in tumorous zona fasciculata tissue may play a role in the pathogenesis of canine cortisol-secreting ATs.     

A comparison of Freund's Complete and Freund's Modified Adjuvants used with a contraceptive vaccine in wild horses (Equus caballus).

Fifteen captive wild mares (Equus caballus) were treated with porcine zona pellucida contraceptive vaccine and either Freund's Complete Adjuvant (n = 7) or Freund's Modified Adjuvant (n = 8). All mares received a booster inoculation of porcine zona pellucida plus Freund's Incomplete Adjuvant a month later. Anti-porcine zona pellucida antibodies were measured over 10 mo following the initial inoculation. There were no significant differences in antibody titers at any point during the 10 mo, and seven of the eight mares in the Freund's Modified Adjuvant group were above the 60% level at the end of the study, which is considered to be the contraceptive threshold for horses. There were no significant differences in titers between pregnant and nonpregnant horses, nor was there a significant correlation between age and titers. One local injection site reaction occurred after booster treatment with Freund's Incomplete Adjuvant, and 11 healthy foals were born during the course of the study. These data suggest that Freund's Modified Adjuvant is an acceptable substitute for Freund's Complete Adjuvant in certain free-ranging and captive wildlife species.     

Comparison of the histological changes in the dog after treatment with the progestins medroxyprogesterone acetate and proligestone

Abstract

Administration of progestins in the dog may result in overproduction of growth hormone, suppression of the hypothalamic‐pituitary‐adrenocortical axis, and insulin resistance. In this paper we present a comparison of the histological findings in control dogs and dogs treated with either medroxyprogesterone acetate (MPA) or proligestone (PROL).

Depot preparations of MPA or PROL were administered (SC) at 3‐week intervals in two groups of seven ovariohysterectomized beagle dogs, after which three dogs of each group were killed. After a 6‐month period without hormone treatment during which recovery was studied, the remaining dogs received five additional injections at the same interval and were subsequently killed. Tissue samples of four intact female beagle dogs served as controls.

Progestin treatment resulted in atrophy of the adrenal cortex. In both MPA‐ and PROL‐treated dogs, the thickness of the combined zona fasciculata and reticularis was significantly smaller than in control animals. In the mammary glands of progestin‐treated dogs there were well developed alveoli and normal ducts adjacent to foci of hyperplastic ductular epithelium. Five dogs in each treatment group had developed benign mammary tumours which varied from simple tubular and papillary adenomas to benign complex and mixed tumours, whereas no mammary tumours were observed in the control animals. In each treatment group, steroid‐induced hepatopathy was observed in the liver of three dogs. Vacuolation of the cells of the islets of Langerhans and the epithelium of the intercalated ducts was present in two dogs of each treatment group and was only observed after the second series of progestin administrations. Incidental findings included chronic pyelonephritis, aspecific dermatitis, and mucinous dysplasia of the gall bladder. No abnormalities were found in sections of spleen, lung, brain, or pituitary gland.

There were no significant differences in the frequencies of the various abnormalities between MPA‐ and PROL‐treated dogs. Our findings correspond with the clinical and biochemical results after treatment of dogs with MPA and PROL. The high incidence of mammary tumours might be associated with our recent finding that in the dog progestins induce ectopic production of growth hormone in the mammary gland. The dog might be a good model for further studies on hormonally induced breast cancers.     

Histological Structure of the Adrenal Gland of the Bottlenose Dolphin (Tursiops truncatus) and the Striped Dolphin (Stenella coeruleoalba) from the Adriatic Sea

The structure of the adrenal gland was studied in 11 bottlenose dolphins ( Tursiops truncatus ), and five striped dolphins ( Stenella coeruleoalba ). These species are legally protected in Croatia. All examined animals died of natural causes and were found stranded along eastern Adriatic coast. In both species the adrenal gland consists of a cortex and a medulla; the cortex is divided into three zones. Whereas in the bottlenose dolphin, there is a zona arcuata which contains columnar cells arranged in the form of arches; in the striped dolphin this zone is replaced by zona glomerulosa containing rounded clusters of polygonal cells. In both species, the zona fasciculata consists of radially oriented cords of polygonal cells, whereas in zona reticularis cells are arranged in branching and anastomosing cords. The adrenal medulla in both species contains dark, epinephrine-secreting cells and light norepinephrine-secreting cells. Epinephrine-secreting cells are localized in the outer part of the medulla, whereas norepinephrine-secreting cells are found in the inner part, arranged in clusters and surrounded by septa of thin connective tissue. The gland is surrounded by a thick connective-tissue capsule, from where thick trabeculae extend towards the interior. In the bottlenose dolphin, group of cells resembling both medullar and cortical cells can be seen within the capsule; whereas only groups of cells resembling cortical cells are found within the capsule of the striped dolphin. In the bottlenose dolphin invagination of the adrenal cortex into the medulla is obvious as well as medullary protrusions extending through cortex to the connective tissue capsule.     

三苯基锡对SD大鼠肾上腺组织结构的影响

[目的]研究环境水平三苯基锡（triphenyltin,TPT）对哺乳动物肾上腺的毒性作用。[方法]将40只青春期SD大鼠随机分为3个不同剂量的试验组[0.5、5、50μg/（kg·BW）]和1个对照组[0μg/（kg·BW）]。每3 d灌胃1次,暴露54 d后,用10%水合氯醛麻醉并处死大鼠,采用石蜡切片和HE染色的方法观察SD大鼠肾上腺组织学变化特点。[结果]对照组SD大鼠肾上腺组织结构正常,皮质部球状带、束状带、网状带内细胞结构正常,胞核明显;髓质部细胞呈多边形或圆形,排列不规则,索间有血窦。与对照组相比,低剂量组肾上腺皮质和髓质内细胞的结构均无明显变化。中剂量组肾上腺球状带稍变薄,束状带和髓质细胞的胞核固缩、胞质溶解,间质组织轻微出血。高剂量组球状带细胞显著减少,与束状带界限不清,束状带细胞和髓质部细胞胞核固缩,胞质溶解现象加重,呈现细胞空泡化现象,间质组织增厚且出血现象明显。[结论]证实了一定剂量的TPT对SD大鼠肾上腺组织结构具有一定损伤作用,且呈剂量依赖性,该结果为研究TPT对哺乳动物内分泌的影响提供了形态学资料。     

Adrenal glands of slaughtered bulls, heifers and cows: a histological study

The study involved histological and immunohistochemical examinations of the adrenal glands of healthy slaughtered cattle. Glands of 13 bulls, 10 heifers and 10 cows were examined. The following histological findings were observed: Unequal thickness of connective capsule and nodular formations of the zona glomerulosa (ZG), eosinophilic granules in cells of the ZG, globoid arrangement of the zona fasciculata, nodules or pegs of cortical tissue in the medulla, mutual interlacing of superficial and deep zones of the medulla, proliferation of cortical or medullary cells into the blood vessels wall situated in the medulla and focal inflammatory infiltrates. Cortical cells and noradrenalin-secreting (N) cells in the medulla expressed cytoplasmic positivity of S100 protein. Both adrenalin (A) cells and N cells were positive in synaptophysin. The majority of the cells in the cortex and in the medulla displayed were positive for chromogranin A. Electron microscopy showed structureless, electrondense particles of varying size and shape, mostly displaying the having mostly character of secretory granules.     

Experimental intraamnionic exposure of bovine fetuses with subgroup 2, type 7 Adenovirus.

Three third trimester fetuses were exposed to a subgroup 2, type 7 adenovirus (adeno 7) by intraamnionic infection. The virus caused preterm delivery of two clinically ill calves and one stillbirth. The two premature calves died 12 and 72 hours after birth. An elevated serum neutralizing antibody titer (1:256) to adenovirus 7 was found in one principal calf at birth. Adenovirus 7 was recovered from several tissues of the live calves and the spleen of the stillborn calf. Fetuses exposed by intraamniotnic injection with virus carrier only, were born healthy after normal gestational periods and no viruses were isolated from the tissues. Clinically ill calves were weak, severely depressed and unable to stand and nurse. Gross postmortem lesions were nonspecific and consisted of petechial and ecchymotic hemorrhages and edema of the gastrointestinal tract. Histopathological lesions included vasculitis, necrosis of the mucosa of the forestomach, mild gastroenteritis and acute, nonsuppurative focal necrosis of the liver, kidney and adrenal gland. Intranuclear inclusion bodies were seen in pericytes, macrophages, hepatocytes, epithelial cells of adrenal cortical sinusoids of the zona glomerulosa and zona fasciculata and renal tubular epithelium.     

Prenatal development of the adrenal gland in the one-humped camel (Camelus dromedarius)

With 14 figures and 3 tables SUMMARY: Each adrenal gland consisted of cortex and medulla that developed from different embryological origins and presented different cellular organization. One hundred male or female camel embryos or fetuses with crown vertebral rump lengths (CVRL) that ranged from 0.8 to 117 cm were examined. The adrenal cortex, which is derived from intermediate mesoderm, was first observed in the 0.8-cm CVRL camel embryo. The adrenal cortex initially was combined with the gonad as a thickened region of proliferating cells derived from splanchnic intermediate mesoderm. Adrenocortical tissue was first separated from the gonadal tissue in the 2-cm CVRL camel fetus and was observed as a separate dorso-medial mass of cells. At 2.5-cm CVRL, the adrenocortical tissue was surrounded by a capsule of undifferentiated mesenchymal cells, except at its proximal pole, where an invagination was located through which chromaffinoblast cells entered the cortex. The chromaffinoblast cells migrated from the neural crest to form the medulla of the developing adrenal gland. In the 3.5-cm CVRL camel fetus, the adrenocortical cells differentiated into two layers: the inner fetal cortex and the outer definitive cortex. As development proceeded, the fetal cortex degenerated and the definitive cortex formed the zona glomerulosa and zona fasciculata. The zona reticularis did not form until the end of gestation. During prenatal life, the adrenal medulla was much thicker than the cortex.     

Developmental competence of in vitro matured ovine oocytes vitrified in solutions with different concentrations of trehalose

This study aimed to determine the optimum concentration of trehalose in solutions used for vitrification of in vitro matured (IVM) ovine oocytes. IVM oocytes were randomly divided into four experimental (vitrified) and one control (fresh) groups. Experimental groups were treated with different concentrations (0.0, 0.25, 0.5 and 1.0 M) of trehalose. After warming, some viable oocytes were exposed to 0.25% pronase to test zona pellucida hardening, whereas the others were fertilized and cultured in vitro for 8 days to evaluate their developmental competence. Blastocysts quality was assessed by differential staining and TUNEL test. Survival and developmental rates of oocytes vitrified in the presence of 0.5 M trehalose were significantly higher than those of the other vitrified groups. Furthermore, there was a significant difference between fresh and vitrified groups in total blastocyst rate. Analysis of blastocysts quality also revealed a significant difference between the group treated with 0.5 M trehalose and other groups in terms of apoptotic index. Furthermore,zona pellucida digestion time period was longer in trehalose‐free (0.0 M) group compared to other groups. In conclusion, we found that IVM ovine oocytes vitrified in solutions containing 0.5 M trehalose are fertilization‐competent and are able to produce good‐quality blastocysts with an apoptotic index comparable to that of the fresh oocytes. Therefore, 0.5 M may be considered the optimum concentration of trehalose to be used in solutions prepared for vitrification of oocytes.