Vacuolar protein sorting 36(VPS36),a protein primarily known for its role in the Endosomal Sorting Complex Required for Transport pathway,has recently been shown to be linked to chicken reproduction.Previous research ...Vacuolar protein sorting 36(VPS36),a protein primarily known for its role in the Endosomal Sorting Complex Required for Transport pathway,has recently been shown to be linked to chicken reproduction.Previous research showed that Vps36 is significantly downregulated in sexually mature chicken ovaries compared to immature ones.In this study,using real-time quantitative RT-PCR,we investigated the expression pattern of Vps36 and its head-to-head gene Ckap2 mRNA in chicken follicles.Small white follicles were found to have significantly higher expression of Vps36 and Ckap2 mRNA than any other sized follicles(P<0.05).The expression of Vps36 and Ckap2 mRNA were detected in both granulosa and theca layers of pre-ovulatory follicles,the expression of Ckap2 in theca layers was slightly higher than in granulosa cells.Treatment of small yellow follicles with folliclestimulating hormone and estradiol resulted in a marked decrease of both Vps36 and Ckap2 mRNA(P<0.05);however,progesterone,transforming growth factor-β1 and luteinizing hormone induced no significant changes in Vps36 and Ckap2 mRNA expression in these follicles.These results indicate that the head-to-head genes of Vps36 and Ckap2 exhibit similar expression in chicken follicles and are involved in chicken follicle development.展开更多
In the bone marrow and spleen,the developing B cell populations undergo both negative and positive selections to shape their B cell receptor repertoire.To gain insight into the shift of the immunoglobulin heavy(IgH)ch...In the bone marrow and spleen,the developing B cell populations undergo both negative and positive selections to shape their B cell receptor repertoire.To gain insight into the shift of the immunoglobulin heavy(IgH)chain repertoire during B cell development,we undertook large scale Igμchain repertoire analysis of pre-B,immature B and spleen B cell populations.We found that the majority of VH gene segments,VH families,JH and D gene segments,were observed to have significantly different usage frequencies when three B cell populations were compared,but the usage profile of the VH,D,and JH genes between different B cell populations showed high correlations.In both productive and nonproductive rearrangements,the length of CDRH3 shortened significantly on average when B cells entered the periphery.However,the CDRH3 length distribution of nonproductive rearrangements did not follow a Gaussian distribution,but decreased successively in the order 3n–2,3n–1 and 3n,suggesting a direct correlation between mRNA stability and CDRH3 length patterns of nonproductive rearrangements.Further analysis of the individual components comprising CDRH3 of productive rearrangements indicated that the decrease in CDRH3 length was largely due to the reduction of N addition at the 5′and 3′junctions.Moreover,with development,the amino acid content of CDRH3 progressed toward fewer positively charged and nonpolar residues but more polar residues.All these data indicated that the expressed Igμchain repertoire,especially the repertoire of CDRH3,was fine-tuned when B cells passed through several checkpoints of selection during the process of maturation.展开更多
Heavy chain only antibodies(HCAbs)represent a rare type of antibody that is devoid of light chains and the CH1 domain that have been reported in cartilaginous fish and camelids.By analyzing transcript data and genome ...Heavy chain only antibodies(HCAbs)represent a rare type of antibody that is devoid of light chains and the CH1 domain that have been reported in cartilaginous fish and camelids.By analyzing transcript data and genome sequences,we conducted a comprehensive analysis of Bactrian camel immunoglobulin heavy chain genes.Based on the transcript data,oneμgene,fiveγgenes,oneαgene and oneεgene were found.Additionally,the variable region of HCAbs(VHH)and the conventional antibodies(VH)sequences associated with theγ3,γ1a/b andμgenes were amplified.Based on these genome sequences,seven DH,six JH,μ,γ2a,γ2c,α,andεgenes and a portion of aγ3 gene were observed.Different Kozak sequences within different VH families were found in our analysis,and the variability index differed between the VHH3 and VH3 families.Phylogenetic analysis of the constant regions of the camelid immunoglobulin genes indicates that these genes appeared before the evolutionary divergence of Bactrian camels and dromedaries.展开更多
基金This work was funded by the National Natural Science Foundation of China(30871777)Platform Construction of Genetic Resources of Livestock and Poultry Breeds in China(2005DKA21101)Agricultural Elite Breeds(Poultry)Project of Shandong Province(2009LZ09-03).
文摘Vacuolar protein sorting 36(VPS36),a protein primarily known for its role in the Endosomal Sorting Complex Required for Transport pathway,has recently been shown to be linked to chicken reproduction.Previous research showed that Vps36 is significantly downregulated in sexually mature chicken ovaries compared to immature ones.In this study,using real-time quantitative RT-PCR,we investigated the expression pattern of Vps36 and its head-to-head gene Ckap2 mRNA in chicken follicles.Small white follicles were found to have significantly higher expression of Vps36 and Ckap2 mRNA than any other sized follicles(P<0.05).The expression of Vps36 and Ckap2 mRNA were detected in both granulosa and theca layers of pre-ovulatory follicles,the expression of Ckap2 in theca layers was slightly higher than in granulosa cells.Treatment of small yellow follicles with folliclestimulating hormone and estradiol resulted in a marked decrease of both Vps36 and Ckap2 mRNA(P<0.05);however,progesterone,transforming growth factor-β1 and luteinizing hormone induced no significant changes in Vps36 and Ckap2 mRNA expression in these follicles.These results indicate that the head-to-head genes of Vps36 and Ckap2 exhibit similar expression in chicken follicles and are involved in chicken follicle development.
基金This work was supported by the National Basic Research Program of China(2010CB945300).
文摘In the bone marrow and spleen,the developing B cell populations undergo both negative and positive selections to shape their B cell receptor repertoire.To gain insight into the shift of the immunoglobulin heavy(IgH)chain repertoire during B cell development,we undertook large scale Igμchain repertoire analysis of pre-B,immature B and spleen B cell populations.We found that the majority of VH gene segments,VH families,JH and D gene segments,were observed to have significantly different usage frequencies when three B cell populations were compared,but the usage profile of the VH,D,and JH genes between different B cell populations showed high correlations.In both productive and nonproductive rearrangements,the length of CDRH3 shortened significantly on average when B cells entered the periphery.However,the CDRH3 length distribution of nonproductive rearrangements did not follow a Gaussian distribution,but decreased successively in the order 3n–2,3n–1 and 3n,suggesting a direct correlation between mRNA stability and CDRH3 length patterns of nonproductive rearrangements.Further analysis of the individual components comprising CDRH3 of productive rearrangements indicated that the decrease in CDRH3 length was largely due to the reduction of N addition at the 5′and 3′junctions.Moreover,with development,the amino acid content of CDRH3 progressed toward fewer positively charged and nonpolar residues but more polar residues.All these data indicated that the expressed Igμchain repertoire,especially the repertoire of CDRH3,was fine-tuned when B cells passed through several checkpoints of selection during the process of maturation.
文摘Heavy chain only antibodies(HCAbs)represent a rare type of antibody that is devoid of light chains and the CH1 domain that have been reported in cartilaginous fish and camelids.By analyzing transcript data and genome sequences,we conducted a comprehensive analysis of Bactrian camel immunoglobulin heavy chain genes.Based on the transcript data,oneμgene,fiveγgenes,oneαgene and oneεgene were found.Additionally,the variable region of HCAbs(VHH)and the conventional antibodies(VH)sequences associated with theγ3,γ1a/b andμgenes were amplified.Based on these genome sequences,seven DH,six JH,μ,γ2a,γ2c,α,andεgenes and a portion of aγ3 gene were observed.Different Kozak sequences within different VH families were found in our analysis,and the variability index differed between the VHH3 and VH3 families.Phylogenetic analysis of the constant regions of the camelid immunoglobulin genes indicates that these genes appeared before the evolutionary divergence of Bactrian camels and dromedaries.
