Characterization of hGFAP-DsRed Transgenic Guangxi Bama Mini-Pigs and Their Offspring
Xiangxing Zhu1, Junyu Nie1, Shouneng Quan1,2, Huiyan Xu1, Xiaogan Yang1, Yangqing Lu1, Kehuan Lu1 and Shengsheng Lu1*
hGFAP-DsRed transgenic Guangxi Bama mini-pig founders showed normal health and development. The hGFAP-DsRed transgenic founders were clinically healthy and developed normally after birth (A), two months old (B) and more than one year old (C). Histological analysis showed that the various brain regions including cerebral cortex (D), olfactory bulb (E), hippocampus (F), substantia nigra (G), cerebellum (H), spinal cord (I) as well as liver (J) and pancreas (K) of the hGFAP-DsRed transgenic founders were normal. (Scale bars = 25 μm).
Astrocyte-specific transgene expression in hGFAP-DsRed transgenic Guangxi Bama mini-pig founders. Among major regions of the central nervous system (CNS) including the cerebral cortex, olfactory bulb, hippocampus as well as the corpus striatum, a large number of bright red fluorescent cells (indicated by arrows) interspersed within the tissues were detected. All of the DsRed-expressing cells exhibited the astrocyte-specific intricate bushy or spongiform morphology, indicating they were astrocytes located in CNS. Conversely, DsRed-expressing cells were absent in the tissues from a wild-type mini-pig as a negative control. (Scale bars = 50 μm).
Germline transgene transmission in offspring. Panel A shows that the offspring (litter No. 2), which were generated by mating hGFAP-DsRed male transgenic founders with wild-type female Guangxi Bama mini-pigs, were clinically healthy and developed normally. The exogenous transgene fragments transmitted in the genome of these offspring were confirmed by PCR genotyping (B). However, the astrocyte-specific expression of DsRed in the transgenic founders was absent among various brain regions including the cortex, olfactory bulb, corpus striatum, hippocampus, cerebellum, and spinal cord, indicating that the inherited transgene was not expressed in the offspring (C). (Scale bars = 100 μm).