TY - JOUR
T1 - The measurement of luteinising hormone in the Western grey kangaroo (Macropus fuliginosis) and black-flanked rock wallaby (Petrogale lateralis lateralis).
AU - Matson, Phillip
AU - Mayberry, Chris
AU - Willers, Nicole
AU - Blackberry, Margaret
AU - Martin, G.B.
PY - 2009
Y1 - 2009
N2 - In the present study, we initially attempted to detect LH in serum of female Western grey kangaroos (Macropus fuliginosis) with a radioimmunoassay (RIA) based on polyclonal antibodies raised in rabbits to four different isoforms of ovine LH: NIH-LH-S11 (3 rabbits), M3-CNRS (from M Jutisz, College de France; 2 rabbits), and oLH-CY-1083 and oLH-CY-1085 (both from Yves Combarnous, INRA, France; 1 rabbit each). None of these antibodies were able to detect any LH in the serum. We then applied an enzymeimmunoassay (EIA) optimised to detect LH in the Asian elephant (Brown et al. 2004) to serum from female M. fuliginosis and black-flanked rock wallabies (Petrogale lateralis lateralis). This assay uses a monoclonal antibody (#518B7) against anti-bovine LH β-subunit (Matteri et al. 1987), provided by Dr Jan Roser (University of California, Davis, USA), biotinylated ovine LH label and bovine LH standard (NIADDK-oLH-26 and NIH-bLH-B10, both provided by Dr Janine Brown and Nicole Abbondanza, Smithsonian Institute, Front Royal, Virginia USA). The antibody shows minimal cross-reaction with FSH from a number of mammalian species (Matteri et al. 1987) and has previously been used in an RIA to detect LH in M. eugenii (McFarlane et al. 1997). Serial dilution of serum down to 1:8 for 7 individual animals of both M. fuliginosis and P. lateralis lateralis showed displacement of the ovine LH label with an average response that was parallel to the bovine standard curve (Fig. 1). Control samples of bovine LH between 1.24-5.30 ng/ml all had between-assay coefficients of variation <9%. Biological validation was achieved by challenging animals with an intramuscular injection of the gonadotrophin-releasing hormone agonist, gonadorelin (GnRHa; Fertagyl®, 2.5 µg/kg) and measuring LH before and 25 minutes after the injection. Gonadorelin is not recognised in the LH assay (Fig. 1). Eight individual female M. fuliginosis were challenged in February 2007, and 19 individual P. lateralis lateralis were challenged with GnRHa in August/September 2007. Statistical analysis using a paired Student’s t-test confirmed significant increases in plasma concentrations of LH after the administration of the GnRHa for both M. fuliginosis (p>0.05) and P. lateralis lateralis (p<0.001) as shown in Figure 2.
AB - In the present study, we initially attempted to detect LH in serum of female Western grey kangaroos (Macropus fuliginosis) with a radioimmunoassay (RIA) based on polyclonal antibodies raised in rabbits to four different isoforms of ovine LH: NIH-LH-S11 (3 rabbits), M3-CNRS (from M Jutisz, College de France; 2 rabbits), and oLH-CY-1083 and oLH-CY-1085 (both from Yves Combarnous, INRA, France; 1 rabbit each). None of these antibodies were able to detect any LH in the serum. We then applied an enzymeimmunoassay (EIA) optimised to detect LH in the Asian elephant (Brown et al. 2004) to serum from female M. fuliginosis and black-flanked rock wallabies (Petrogale lateralis lateralis). This assay uses a monoclonal antibody (#518B7) against anti-bovine LH β-subunit (Matteri et al. 1987), provided by Dr Jan Roser (University of California, Davis, USA), biotinylated ovine LH label and bovine LH standard (NIADDK-oLH-26 and NIH-bLH-B10, both provided by Dr Janine Brown and Nicole Abbondanza, Smithsonian Institute, Front Royal, Virginia USA). The antibody shows minimal cross-reaction with FSH from a number of mammalian species (Matteri et al. 1987) and has previously been used in an RIA to detect LH in M. eugenii (McFarlane et al. 1997). Serial dilution of serum down to 1:8 for 7 individual animals of both M. fuliginosis and P. lateralis lateralis showed displacement of the ovine LH label with an average response that was parallel to the bovine standard curve (Fig. 1). Control samples of bovine LH between 1.24-5.30 ng/ml all had between-assay coefficients of variation <9%. Biological validation was achieved by challenging animals with an intramuscular injection of the gonadotrophin-releasing hormone agonist, gonadorelin (GnRHa; Fertagyl®, 2.5 µg/kg) and measuring LH before and 25 minutes after the injection. Gonadorelin is not recognised in the LH assay (Fig. 1). Eight individual female M. fuliginosis were challenged in February 2007, and 19 individual P. lateralis lateralis were challenged with GnRHa in August/September 2007. Statistical analysis using a paired Student’s t-test confirmed significant increases in plasma concentrations of LH after the administration of the GnRHa for both M. fuliginosis (p>0.05) and P. lateralis lateralis (p<0.001) as shown in Figure 2.
U2 - 10.1071/AM08112
DO - 10.1071/AM08112
M3 - Article
SN - 0310-0049
VL - 31
SP - 61
EP - 63
JO - Australian Mammalogy
JF - Australian Mammalogy
IS - 1
ER -