Gender-dependent physiological implications of combined PAI-1 and TIMP-1 gene deficiency characterized in a mouse model

Abstract

The endogenous proteinase inhibitors plasminogen activator inhibitor type 1 (PAI-1) and tissue inhibitor of metalloproteinase type 1 (TIMP-1) are two distinct proteins with separate molecular pathways. However, a close relationship between PAI-1 and TIMP-1 has been proposed indicating some degree of functional overlap due to their involvement in ECM turnover, tissue remodeling, and cellular migration and signaling. To study the housekeeping physiological implications of PAI-1 and TIMP-1, we generated a combined PAI-1 and TIMP-1 gene-deficient mouse model. We present the results on generating this specific mouse model with particular emphasis on phenotypical characteristics, blood leukocyte counts, histology, and gene expression studies of PAI-1 and TIMP-1 in various organs. We observed a significant deviation in segregation of offspring only in male mice (P < 0.01) predominantly caused by PAI-1 deficiency. In addition, the body weight in 3- and 20-wk-old male and 20-wk-old female mice was significantly different between genotypes (P <or= 0.0008). Furthermore, blood leukocyte counts were significantly different between genotypes in 20-wk-old male mice (P <or= 0.0002), whereas no significant differences were observed between genotypes in 20-wk-old female mice (P >or= 0.13). Quantifying the relative expression of PAI-1 and TIMP-1 revealed upregulation of PAI-1 (P < 0.001) in male mice only. Our data highlight the complex roles of PAI-1 and TIMP-1 on physiological parameters such as segregation of offspring (embryonic development and survival), body weight (metabolism), blood leukocyte counts (immunity), and gene expression (regulatory redundancy). We conclude that PAI-1 and TIMP-1 seem to possess gender-dependent regulatory properties on various housekeeping physiological parameters and stress the potential implications in pathological conditions.

Loss of huntingtin-associated protein 1 impairs insulin secretion from pancreatic β-cells

“…2c). Although the mechanism for the sex-dependent difference in body weight, which also occurs in mice null for other genes [39], remains to be investigated, depletion of Hap1 in β -cells in the pancreas seems to have more impact on the body weight of male mice.…”

NGF Upregulates the Plasminogen Activation Inhibitor-1 in Neurons via the Calcineurin/NFAT Pathway and the Down Syndrome-Related Proteins DYRK1A and RCAN1 Attenuate This Effect

“…For cDNA synthesis 1 µg of total RNA was reverse transcribed using 1 µg of oligo(dT) primers and 2000 units of MMLV reverse transcriptase (Promega) at 40°C for 1 h. The resulting cDNAs were analysed using a LightCycler 480 system and SYBR Green master mix reagent (Roche Applied Science) using the following PCR conditions: 5 min initial denaturation at 95°C, followed by 45 cycles of 10 s at 95°C, 10 s at 50-60°C, 15 s at 72°C and 1 s at 74°C. The following oligonucleotide primers were used for the detection of the rat PAI-1 transcript: rPAI-1 for : 5′-GAGGATGAAAGAAACAGCCAGCT-3′ and rPAI-1 rev : 5′-CCCGCTATGAAATTAGATTCACGT-3′ [70], for the detection of the murine PAI-1 transcript: mPAI-1for: 5′-GTACTGCGGATGCCATCTTT-3′ and mPAI-1rev: 5′-GCGTGTCAGCTCGTCTACAG-3′ [71] and for the detection of the rat RCAN1-4 transcript: RCAN1-4 for : 5′-GCCCGTTGAAAAAGCAGAAT-3′ and RCAN1-4 rev : 5′- GACAGGGGGTTGCTGAAGTT-3′ [37]. The beta-2 microglobulin transcript was used as a housekeeping gene for normalization.…”

Lymphotropism and host responses during acute wild-type canine distemper virus infections in a highly susceptible natural host

“…Populations of granulocytes, lymphocytes and monocytes were gated based on their light-scatter characteristics in a forward scatter versus side scatter diagram as described previously (Jensen et al, 2003). For determining absolute counts of leukocytes, the instrument particle (cell or latex bead) registration efficiency was measured by using BD TruCount tubes (Becton Dickinson) (Harslund et al, 2007). The staining methods for subpopulations of mink PBMCs using cell-surface markers have been described previously (Castelruiz et al, 2005;Chen et al, 1997;Jensen et al, 2003).…”