Ultrastructural Morphometry of Mammary Gland in Transgenic and Non‐transgenic Rabbits

Dragin, S.; Pivko, J.; Massányi, Peter; Lukáč, Norbert; Makarevich, Alexander V.; Paleyanda, Rekha K.; Chrenek, Peter

doi:10.1111/j.1439-0264.2006.00692.x

Cited by 7 publications

(6 citation statements)

References 28 publications

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“…All transgenic rabbit females of F 1 , F 2 and F 3 generation had normal reproductive behaviour, producing litters ranging from 7 to 9 offspring, which is in accordance with our previous results (Chrenek et al 2005). The transgenic animals did not exhibit pathological signs such as infertility, respiratory distress or impairment of mammary gland development and lactation, compared to non-transgenic rabbits, as it was previously found (Suvegova et al 2004;Dragin et al 2006). In this study we showed that there were no significant differences between transgenic and non-transgenic females in milk yield investigated on day 10, 21 and 30 at the first lactation.…”

Section: Discussionsupporting

confidence: 94%

Expression of recombinant human factor VIII in milk of several generations of transgenic rabbits

Chrenek

Ryban

Vetr³

et al. 2007

Transgenic Res

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Transgenic founder rabbits carrying a gene construct consisting of a 2.5 kb murine whey acidic protein promoter (mWAP), 7.2 kb of the human clotting factor VIII (hFVIII) cDNA and 4.6 kb of 3' flanking sequences of mWAP gene were crossed for three generations. All transgenic animals showed stable transgene transmission. Transgenic females showed high level of recombinant hFVIII (rhFVIII) mRNA expression in biopsed mammary gland tissues, while marginal expression of rhFVIII mRNA was observed in the spleen, lung and brain. No adverse effects of ectopic expression on the physiology of the rabbits were observed. Expression was not detected in the liver, kidney, heart and skeletal muscle. In transgenic females derived from three generations, rhFVIII protein was secreted from the mammary gland of lactating females, as shown by Western blotting. Biological activity of rhFVIII protein, as revealed in clotting assays was ranged from 0.012 to 0.599 IU/ml corresponding to 1.2% and 59.9% of the hFVIII level in normal human plasma. No apparent effect of secreted rhFVIII on the milk performance of rabbits was observed. Our results confirm the possibility of producing a significant amount of a biologically active rhFVIII in the mammary gland of established transgenic rabbit lines.

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Section: Discussionsupporting

confidence: 94%

Expression of recombinant human factor VIII in milk of several generations of transgenic rabbits

Chrenek

Ryban

Vetr³

et al. 2007

Transgenic Res

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“…Our RT-PCR results showed that high mWAP-hFVIII expression without any deleterious effect was observed in the mammary gland (Dragin et al, 2006), whilst low-level (background) gene expression was detected in the spleen and the lung. Additionally, we compared haematological and biochemical parameters between transgenic and non-transgenic rabbits.…”

Section: Discussionmentioning

confidence: 70%

“…The samples were fixed in 10% formalin, dehydrated in a graded series of ethanol, embedded in paraffin wax and sectioned on a microtome. The serial 10 μm thick sections [five blocks and 10 sections of each organ (total 50 sections per organ)] were stained with haematoxylin and eosin (Dragin et al., 2006).…”

Section: Methodsmentioning

confidence: 99%

Evaluation of Haematological, Biochemical and Histopathological Parameters of Transgenic Rabbits

Jurčík¹,

Süvegová²,

Hanusová³

et al. 2007

Journal of Veterinary Medicine Series A

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The aim of our study was to compare the hFVIII mRNA expression in different organs, pathological changes and selected haematological and biochemical blood parameters between transgenic and non-transgenic rabbits from F3 generation. Selected physiological parameters of 3- to 4-month-old transgenic rabbits from F3 generation carrying human factor VIII gene (hFVIII) were analysed and compared with those of non-transgenic ones. Before slaughtering, the blood for haematological and biochemical analysis was taken from the central ear artery. Pathological and histological examination of vital organs and RT-PCR analysis of several tissue organs of transgenic and non-transgenic animals were performed after slaughtering. Except for the mammary gland tissue, slight hfVIII mRNA expression in the spleen, lung and brain and none expression in the liver, kidney, skeletal muscle and heart of rabbits were recorded. pathological examination of vital organs showed some pathological changes in both transgenic and non-transgenic rabbits which were confirmed by histological qualitative evaluations. Statistically significant lower values of blood haemoglobin in blood of transgenic (11.86+/-0.86) animals compared with non-transgenic (12.41+/-1.02, P<0.05) ones and lower parameters of HCT (39.22+/-2.44 versus 40.89+/-2.26, P<0.01) in blood of transgenic rabbits were observed. Parameters of WBC, RBC and PLT showed no significant differences between the analysed groups. All biochemical serum parameters of transgenic rabbits were higher in comparison with non-transgenic ones. Significant differences were found in the concentration of the urea, AST and GMT between transgenic and non-transgenic animals (P<0.001) and in the total protein content, the difference was significant at P<0.05. In conclusion, our results showed that no considerable impact on the general health was found in transgenic rabbits.

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“…Our recent histological analyses of mammary gland tissues of lactating transgenic rabbit (carrying hFVIII transgene) showed no changes, when compared to nontransgenic mammary gland tissues (Dragin et al, 2006).…”

Section: Discussionmentioning

confidence: 73%

The Yield and Composition of Milk from Transgenic Rabbits

Chrenek¹,

Chrastinová²,

Kirchnerová³

et al. 2007

Asian Australas. J. Anim. Sci

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Basic objective of this research was to compare the milk yield and composition of New Zealand White transgenic rabbit females expressing recombinant human factor VIII (hFVIII) in mammary gland during lactation with that of non-transgenic rabbit females of the same age during 30 days of lactation. Transgenic founders were generated by the microinjection of foreign DNA (mWAP-hFVIII gene construct) into the egg. F1, F2 and F3 generations of transgenic rabbits were obtained after mating of transgenic founder rabbits with non-transgenic rabbits. The amount of milk rejected was measured by weight-suckle-weight method at 10 th , 20 th and 30 th day of lactation. Quality of milk (content of fat, protein, lactose, dry ash, and some minerals) from transgenic and non-transgenic rabbit was also determined. Comparison of milk yield, determined by weight-suckle-weight method, showed significantly higher (p<0.05) milk production at day 20 of first lactation in non-transgenic females, but on the same day of second lactation higher milk yield was measured in transgenic ones. Significantly higher (p<0.05) content of milk fat and protein was determined in transgenic milk whilst higher content of lactose was found in non-transgenic milk. The content of minerals (calcium, phosphorus, magnesium and sodium) did not differ in both experimental and control groups. Our results showed that milk yield and composition of transgenic rabbit females (mammary specific transgenic over-expression of hFVIII) over several generations is only slightly and transiently different from milk yield of nontransgenic females, which had no significant consequence on the litter size and viability.

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Ultrastructural Morphometry of Mammary Gland in Transgenic and Non‐transgenic Rabbits

Cited by 7 publications

References 28 publications

Expression of recombinant human factor VIII in milk of several generations of transgenic rabbits

Expression of recombinant human factor VIII in milk of several generations of transgenic rabbits

Evaluation of Haematological, Biochemical and Histopathological Parameters of Transgenic Rabbits

The Yield and Composition of Milk from Transgenic Rabbits

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