Exogenous DNA internalisation by sperm cells is improved by combining lipofection and restriction enzyme mediated integration

1. Three types of exogenous DNA inserts, i.e. complete linearised pVIVO2-GFP/LacZ vector (9620 bp), the LacZ gene (5317 bp) and the GFP gene (2152 bp) were used to transfect chicken spermatozoa through simple incubation of sperm cells with insert. 2. PCR assay, Dot Blot hybridisation and Southern hybridisation showed the successful internalisation of exogenous DNA by chicken sperm cells. 3. Lipofection and Restriction Enzyme Mediated Integration (REMI) were used to improve the rate of internalisation of exogenous DNA by sperm cells. 4. Results from dot blot as well as Southern hybridisation were semi-quantified and improved exogenous DNA uptake by sperm cells through lipofection and REMI. Stronger signals were observed from hybridisation of LacZ as well as GFP specific probe with the DNA from lipofected exogenous DNA transfected sperm DNA in comparison with those transfected with nude exogenous DNA.     

Toxicological effects and recovery of the corneal epithelium in Cyprinus carpio communis Linn. exposed to monocrotophos: an scanning electron microscope study

This study was conducted based on the evidence of fish habitats in North India being affected by organophosphate pesticides draining from agricultural fields into bodies of water, especially during the rainy season. Various tissues of fish such as scales, gills ovaries, kidney, and liver have been studied from the toxicological point of view, but the toxicological effects of aquatic pollutants on fish cornea have not been investigated to date. We conducted comparative toxicological studies on the cornea of Cyprinus carpio communis using two sublethal (0.038 and 0.126 ppm) concentrations of monocrotophos pesticide for 30 days. Corneas from all the groups were evaluated by a scanning electron microscope. The fish exposed to the monocrotophos pesticide developed corneal necrosis due to the formation of crystalloid‐like structures, thinning and shrinkage of microridges on the corneal epithelium. After 30 days, fish from the monocrotophos‐treated tank were transferred to normal environmental conditions. After 60 days under natural condition, epithelial cells did not fully recover. In conclusion, exposure to monocrotophos induces irreversible changes in the cornea of C. carpio communis. As fish and mammalian visual systems share many similarities, the reported finding may offer useful insights for further toxicological and ophthalmological studies in humans.     

ARTERIOGRAPHIC ANATOMY OF THE ABDOMINAL AORTA IN THE GOAT, DOG, PIG, AND RABBIT

Arteriographic anatomy of the major branches of the abdominal aorta of rabbits, dogs, pigs, and goats was investigated. The origin and location of the celiac, cranial mesenteric, right renal, left renal, caudal mesenteric, and external iliac arteries were demonstrated, and their placement was described in relation to the iliac bifurcation and lumbar vertebral bodies.     

Uterine environment as a regulator of birth weight and body dimensions of newborn lambs

Pure-bred embryos were transferred within and reciprocally between large (Suffolk) and small (Cheviot) breeds of sheep to establish 4 treatment groups: SinS (Suffolk embryos in Suffolk dams), SinC (Suffolk embryos in Cheviot dams), CinS (Cheviot embryos in Suffolk dams), and CinC (Cheviot embryos in Cheviot dams). The recipient ewes carried single fetuses to term. The maternal plasma concentrations of ovine placental lactogen (oPL), progesterone, IGF-1, FFA, and glucose were measured on d 50, 90, 120, and 140 of pregnancy. Birth weight, body dimensions, and placental characteristics of lambs were recorded at birth. There was a recipient ewe breed × lamb breed × time interaction for the concentration of oPL (P = 0.03), but no such interaction was observed for progesterone (P = 0.42), IGF-1 (P = 0.57), glucose (P = 0.36), or FFA (P = 0.72). There were no differences in oPL (P = 0.28) and progesterone (P = 0.34) concentrations between SinC and SinS ewes. The concentrations of FFA on d 140 (P = 0.008), and those of glucose on d 50 (P = 0.02) and 120 (P = 0.01), were greater in SinC ewes than in SinS ewes. The ewes in CinS had less FFA concentration (P = 0.002) at all time points than CinC ewes. The concentrations of IGF-1 on d 90 were greater (P = 0.004) in CinS ewes than CinC ewes, but did not differ (P = 0.16) on d 50, 120, and 140. The concentrations of glucose on d 50 (P = 0.001), 90 (P = 0.03), and 140 (P = 0.03) were less in CinS ewes compared with CinC ewes. The birth weight of SinC lambs (5.04 ± 0.20 kg) was lighter (P = 0.001) than SinS lambs (5.94 ± 0.19 kg), and body dimensions of SinC lambs were smaller (P = 0.01) than SinS lambs. Neither birth weight nor the body dimensions of CinS lambs differed (P = 0.24) from CinC lambs. Cotyledon number was reduced (P = 0.04) in the CinS (57.5 ± 6.3) compared with the SinS group (74.2 ± 5.9), whereas mean cotyledon weight in CinS (2.42 ± 0.20 g) was greater (P = 0.02) than SinS (1.74 ± 0.21 g). It was concluded that the large genotype lambs were lighter and smaller when born to small genotype dams; however, the birth weight or body dimensions of small genotype lambs did not differ when born to large genotype dams. This study suggests that plasma oPL, progesterone, IGF-1, FFA, and glucose concentrations at different times throughout pregnancy reflect the regulatory effect of the uterine environment on the development of the fetus.     

Vernalization of immature embryos of winter wheat genotypes

Summary The effect of direct vernalization of immature embryos on flowering was studied in six winter wheat genotypes. Fourteen-, 17-, and 20-day-old embryos were excised and vernalized for 0–6 weeks on synthetic medium during a conditioning period. Percent germination of embryos was high (overall 96.1%), and free from genotypic effects. Genotypes differed for flowering in response to cold treatment of excised embryos. Embryo vernalization was as effective as or more than conventional vernalization (control, seedling vernalization for 6 weeks). Seventeen-day-old embryos were the most responsive to vernalization. With a 5-week vernalization of 17-day-old embryos, the percentage of plants anthesed was higher than those from 14-and 20-day-old embryos. For 17-day-old embryos vernalized for 5 weeks, the mean number of days from culture to anthesis was less than that of 6 week vernalization, less than that of 14- and 20-day-old embryos, and less than controls.Purdue Univ., Agronomy Dept., W. Lafayette, IN 47907, USA.