The importance of genetic research in zoo breeding programmes for threatened species: the African dwarf crocodiles (genus Osteolaemus

The threatened African dwarf crocodiles (genus Osteolaemus) are distributed throughout West and Central Africa. Traditionally two subspecies were described (Osteolaemus tetraspis tetraspis and Osteolaemus tetraspis osborni), although recent molecular studies demonstrate the presence of three allopatric lineages that should be recognized as full species. These highly divergent taxa are distributed in the three major forested biogeographic zones of western Africa: Congolian (Osteolaemus osborni), Lower Guinean (Osteolaemus tetraspis) and Upper Guinean (Osteolaemus sp. nov. cf. tetraspis). Largely because of their diminutive size, dwarf crocodiles are regularly kept in zoos and aquariums worldwide. In Europe, the collection is managed by a European studbook coordinated by Leipzig Zoo, Germany, since 2006, while American zoological institutions do not yet manage these species as part of a studbook programme. To facilitate ex situ conservation efforts, it is important to identify accurately each individual to the appropriate species following the latest systematic understanding of the genus. Population aggregation analysis with mitochondrial and nuclear gene sequences was used for both species identification and detection of interspecific hybridization. The results of our study show that only European collections house all three Osteolaemus taxa, although only a single individual O.osborni was confirmed. The most prevelant species present in both European and North American institutions was O.tetraspis. Additionally, several O.sp. nov. cf. tetraspis were identified, likely originating from the Senegambia region, especially in the North American collections. This will represent an important resource for future conservation efforts as Osteolaemus are highly threatened in this region of West Africa. Unfortunately, both zoo populations showed relatively high frequencies (c. 25-28%) of hybridization between O.tetraspis and O.sp. nov. cf. tetraspis bred in captivity. We highly recommend that zoological institutions ensure they know the species identity of the Osteolaemus they maintain and work together to transfer individuals into single-species colonies to avoid further hybridization. In the USA, this may necessitate the creation of a studbook programme. It may also prove valuable to consider a cooperative programme between the European Association of Zoos and Aquaria and the Association of Zoos & Aquariums, wherein each Association focuses its resources largely on a single Osteolaemus species. This would, however, require trans-Atlantic transfer of individuals. The case study of dwarf crocodiles in zoological institutions reinforces the importance of genetic research in conservation-breeding programmes, highlights the potential for collaboration between European and American zoological institutions for the ex situ conservation of threatened wildlife, and foreshadows some of the regulatory challenges in managing captive populations internationally

Identification of the Penta-EF-hand Protein ALG-2 as a Ca2+-dependent Interactor of Mucolipin-1*

Loss of function mutations in mucolipin-1 (MCOLN1) have been linked to mucolipidosis type IV (MLIV), a recessive lysosomal storage disease characterized by severe neurological and ophthalmological abnormalities. MCOLN1 is an ion channel that regulates membrane transport along the endolysosomal pathway. It has been suggested that MCOLN1 participates in several Ca2+-dependent processes, including fusion of lysosomes with the plasma membrane, fusion of late endosomes and autophagosomes with lysosomes, and lysosomal biogenesis. Here, we searched for proteins that interact with MCOLN1 in a Ca2+-dependent manner. We found that the penta-EF-hand protein ALG-2 binds to the NH-terminal cytosolic tail of MCOLN1. The interaction is direct, strictly dependent on Ca2+, and mediated by a patch of charged and hydrophobic residues located between MCOLN1 residues 37 and 49. We further show that MCOLN1 and ALG-2 co-localize to enlarged endosomes induced by overexpression of an ATPase-defective dominant-negative form of Vps4B (Vps4BE235Q). In agreement with the proposed role of MCOLN1 in the regulation of fusion/fission events, we found that overexpression of MCOLN1 caused accumulation of enlarged, aberrant endosomes that contain both early and late endosome markers. Interestingly, aggregation of abnormal endosomes was greatly reduced when the ALG-2-binding domain in MCOLN1 was mutated, suggesting that ALG-2 regulates MCOLN1 function. Overall, our data provide new insight into the molecular mechanisms that regulate MCOLN1 activity. We propose that ALG-2 acts as a Ca2+ sensor that modulates the function of MCOLN1 along the late endosomal-lysosomal pathway