[Detection of murine tissues that express opticin]

The LRR [leucine rich proteoglycans] is a molecular recognition motif found in proteins with some roles in cell adhesion, signal transduction, DNA repair and RNA processing. Opticin is a member of this family. Takanosu et al [2001] detected messenger RNA expression of mouse opticin in the eye, heart, brain, testis, thyroid and epididymis by dot blot hybridization. In this study, expression levels of mRNA and protein of opticin was investigated by two monoclonal antibodies which were raised against opticin peptides. By this method structure of opticin in human and mouse has been studied. Mouse tissues including, kidney, testis, liver, lung, heart, brain, muscle, spleen and eye were isolated. Opticin expression was identified at mRNA and protein levels by RTPCR and Western blot. PCR analysis revealed that opticin mRNA is expressed in all the tissues studied except for the lung. However, opticin protein was detected in all tissues analyzed. Expression of opticin in the adult murine tissues may suggest functions other than that of putative regulation of vitreous collagen fibrillogenesis for this molecule

Indoleamine 2,3-dioxygenase [IDO] is expressed at feto-placental unit throughout mouse gestation: An immunohistochemical study

The cells expressing Indoleamine 2, 3-dioxygenase [IDO] in feto-maternal interface mediate tryptophan catabolism, hence protect allogeneic fetus from lethal rejection by maternal immune responses. In this study, we report immuno-localization of IDO cells in murine reproductive tract and placenta throughout mouse pregnancy by immunohistochemistry. Syngeneic pregnant mice were examined for vaginal plug to discover about their state of pregnancy. A total of three pregnant mice were examined at each stage. The examination was further confirmed by the detection of sperm in vaginal smear. On the gestational days of 2[nd], 12[th] and 18[th], the uterus and oviduct were removed and expression of IDO was investigated in the endometrium, placenta and oviduct by immunohistochemistry. Our results showed that IDO is expressed consistently in feto-maternal interface throughout pregnancy. In endometrium, expression of IDO was predomin-antly confined to luminal and glandular epithelial cells. Cells at junctional and labyrinth zones of placenta showed strong IDO immunoreactivity as well. Expression of IDO at the protein level in reproductive tract of pregnant mice during entire periods of gestation points to its potential protective role in maintenance of pregnancy. In our knowledge this is the first report of expression of IDO in feto-maternal phase during murine pregnancy

Producing recombinant mTEX101; a murine testis specific protein

Production of antibodies against specific proteins of testis germ cells is of great significance for the investigation of processes involved in spermatogenesis, study of infertility problems and determination of the probable role of these proteins as cancer-testis antigens. Murine Testis Specific Recombinant Protein 101 [mTEX101] is a 38kDa, GPI-anchored protein which is expressed in testis germ cells of adult mice but it seems to be absent in other tissues. The structure and function of mTEX101 is not completely understood yet, but it is speculated that it may transduce biochemical signals into the cytoplasm since mTEX101 does not have an intracellular domain but the precise mechanisms are still ambiguous. RNA was extracted from three adult mice testis. The RNA was used in RT-PCR, employing a pair of specific primers for mTEX101 ORF region. TA-cloning technique was performed by the insertion of mTEX101 into a pGEM-T Easy Vector, followed by its subcloning into a His-tagged expression vector, pET-28a [+]. The recombinant mTEX101 was then produced by transfection of the expression vector into BL 21 [DE3] E. coli strain. A recombinant protein, weighing 27kDa, was produced upon IPTG-induction of the bacterial host. The presence of mTEX101 protein was detected through Western blot analysis by anti-mTEX101 peptide antibodies. We produced mTEX101 recombinant protein that could be used for the production of mono and polyclonal antibodies