Fine structure of the hypobranchial gland of Haliotis diversicolor has been studied by light and electron microscopy. The results showed that the gland is folded pleats organ, which is highly glandular area of the epi...Fine structure of the hypobranchial gland of Haliotis diversicolor has been studied by light and electron microscopy. The results showed that the gland is folded pleats organ, which is highly glandular area of the epidermal lining the roof of the mantle cavity. Two such glandular areas, one on each side of the rectum, occur in H. diversicolor. Left one is much larger than fight one. By light microscope, on H-E stained section, four cell types can be divided： cells with weak basophilic fibrillar elements; with acidophilic granular substance; with strong basophiIic fibrillar elements and ciliated ceils. In the basal lamina region under the gland epithelium, there are a few connective tissues. Surface view of the gland could be seen by scanning electron microscope, there are cilia and different kinds of secretions distributed. By transmission electron microscope, supporting cells, sensory cells and seven types gland cells were observed to form the glandular epithelium. Cells are rich in rough endoplasmic reticulum, smooth muscle fiber and nerve endings were found beneath glandular epithelium, between basal lamina.展开更多
Quantitative real-time reverse transcription-polymerase chain reaction （qRT-PCR） is widely used in studies of gene expression. In most of these studies, housekeeping genes are used as internal references without val...Quantitative real-time reverse transcription-polymerase chain reaction （qRT-PCR） is widely used in studies of gene expression. In most of these studies, housekeeping genes are used as internal references without validation. To identify appropriate reference genes for qRT-PCR in Pacific abalone Haliotis discus hannai, we examined the transcription stability of six housekeeping genes in abalone tissues in the presence and absence of bacterial infection. For this purpose, abalone were infected with the bacterial pathogen Fibrio anguillarum for 12 h and 48 h. The mRNA levels of the housekeeping genes in five tissues （digestive glands, foot muscle, gill, hemocyte, and mantle） were determined by qRT-PCR. The PCR data was subsequently analyzed with the geNorm and NormFinder algorithms. The results show that in the absence of bacterial infection, elongation factor-l-alpha and beta-actin were the most stably expressed genes in all tissues, and thus are suitable as cross-tissue type normalization factors. However, we did not identify any universal reference genes post infection because the most stable genes varied between tissue types. Furthermore, for most tissues, the optimal reference genes identified by both algorithms at 12 h and 48 h post-infection differed. These results indicate that bacterial infection induced significant changes in the expression of abalone housekeeping genes in a manner that is dependent on tissue type and duration of infection. As a result, different normalization factors must be used for different tissues at different infection points.展开更多
基金Acknowledgments： This study was founded by Natural Science Foundation of Guangdong （No. 032248） and Large Instrument Using Fund of South China Agricultural University （ No. 2007Y002 ） .
文摘Fine structure of the hypobranchial gland of Haliotis diversicolor has been studied by light and electron microscopy. The results showed that the gland is folded pleats organ, which is highly glandular area of the epidermal lining the roof of the mantle cavity. Two such glandular areas, one on each side of the rectum, occur in H. diversicolor. Left one is much larger than fight one. By light microscope, on H-E stained section, four cell types can be divided： cells with weak basophilic fibrillar elements; with acidophilic granular substance; with strong basophiIic fibrillar elements and ciliated ceils. In the basal lamina region under the gland epithelium, there are a few connective tissues. Surface view of the gland could be seen by scanning electron microscope, there are cilia and different kinds of secretions distributed. By transmission electron microscope, supporting cells, sensory cells and seven types gland cells were observed to form the glandular epithelium. Cells are rich in rough endoplasmic reticulum, smooth muscle fiber and nerve endings were found beneath glandular epithelium, between basal lamina.
基金Supported by the Knowledge Innovation Program of Chinese Academy of Sciences(No.KSCX2-EW-G-12B)the Knowledge Innovation Program of the Chinese Academy of Sciences(No.KZCX2-EW-Q213)the National High Technology Research and Development Program of China (863 Program)(No.2012AA10A412)
文摘Quantitative real-time reverse transcription-polymerase chain reaction （qRT-PCR） is widely used in studies of gene expression. In most of these studies, housekeeping genes are used as internal references without validation. To identify appropriate reference genes for qRT-PCR in Pacific abalone Haliotis discus hannai, we examined the transcription stability of six housekeeping genes in abalone tissues in the presence and absence of bacterial infection. For this purpose, abalone were infected with the bacterial pathogen Fibrio anguillarum for 12 h and 48 h. The mRNA levels of the housekeeping genes in five tissues （digestive glands, foot muscle, gill, hemocyte, and mantle） were determined by qRT-PCR. The PCR data was subsequently analyzed with the geNorm and NormFinder algorithms. The results show that in the absence of bacterial infection, elongation factor-l-alpha and beta-actin were the most stably expressed genes in all tissues, and thus are suitable as cross-tissue type normalization factors. However, we did not identify any universal reference genes post infection because the most stable genes varied between tissue types. Furthermore, for most tissues, the optimal reference genes identified by both algorithms at 12 h and 48 h post-infection differed. These results indicate that bacterial infection induced significant changes in the expression of abalone housekeeping genes in a manner that is dependent on tissue type and duration of infection. As a result, different normalization factors must be used for different tissues at different infection points.
