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We highlight a novel molecular clock calibration system based on geologically dated river reversal and river capture events. Changes in drainage pattern may effect vicariant isolation of freshwater taxa, and thus provide a predictive framework for associated phylogeographic study. As a case in point, New Zealand's Pelorus and Kaituna rivers became geologically isolated from the larger Wairau River system 70 to 130 kyr BP. We conducted mitochondrial DNA phylogeographic analyses of two unrelated freshwater-limited fish taxa native to these river systems (Gobiomorphus breviceps, n = 63; Galaxias divergens, n = 95). Phylogenetic analysis of combined control region and cytochrome b sequences yielded reciprocally monophyletic clades of Pelorus-Kaituna and Wairau haplotypes for each species. Calibrated rates of molecular change based on this freshwater vicariant event are substantially faster than traditionally accepted rates for fishes but consistent with other recent inferences based on geologically young calibration points. A survey of freshwater phylogeographic literature reveals numerous examples in which the ages of recent evolutionary events may have been substantially overestimated through the use of "accepted" calibrations. We recommend that--wherever possible--biologists should start to reassess the conclusions of such studies by using more appropriate molecular calibrations derived from recent geological events.

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The endothelium-specific antigen PAL-E is expressed in capillaries and veins throughout the body with the exception of the brain, where the antigen is absent from anatomical sites with a patent blood-brain barrier. In this study we determined vascular endothelial staining for PAL-E in the normal eye in relation to the ocular blood-tissue barriers. Immunohistochemical staining of frozen tissue sections of eyes from 22 cornea donors and a number of normal animal autopsy eyes was performed for the PAL-E antigen and the blood-brain barrier marker glucose transporter l. In normal human and animal eyes, endothelial PAL-E staining was absent from the microvasculature in iris, ciliary muscle, optic nerve and retina. In a few normal human eyes, some weakly stained capillaries were observed in the retina and nerve fiber layer, mostly in the peripapillary area. Marked staining of capillaries and venules with PAL-E was observed in the conjunctiva, episclera, sclera, ciliary processes, choriocapillaris and optic nerve head. In general, the endothelial antigen PAL-E is absent from microvessels involved in the blood-ocular and the blood-retinal barriers. PAL-E may therefore be a useful marker to identify pathological breakdown of blood-ocular barriers.

Aim Historical connectivity between drainages may represent an underappreciated contributor to the biodiversity within a catchment. First, we tested whether an isolated population of freshwater-limited galaxiids represents a contribution by past interdrainage connectivity to this system's biodiversity, rather than reflecting intracatchment dispersal or speciation. Second, we sought to distinguish between possible geomorphic processes that could have allowed any past connectivity.Location The Clutha and adjacent Southland region drainages on the South Island of New Zealand.Methods We estimated the phylogeographical relationships among 466 cytochrome b sequences from 'roundhead' Galaxias populations throughout the Southland region and the Clutha drainage, using three different methods. Subsequently, possible geomorphic mechanisms of interdrainage connectivity were examined concurrently with genetic population divergence time estimates obtained by coalescent analysis.Results All samples obtained in the lower Clutha system were genetically attributable to G. gollumoides and reflected at least a third, independent colonization event from Southland involving this species, rather than intradrainage dispersal or speciation by pre-existing Clutha populations. Geomorphological inferences and divergence time estimates of less than 7 ka are consistent with headwater connections across low divides between the lower Clutha and the Catlins region of Southland, rather than anastomosing of main channels during low sea stands. Main conclusionsThe observation of at least three interdrainage connectivity events influencing the diversity and distribution of G. gollumoides lineages within the Clutha system highlights the potential significance of these processes as contributors to the magnitude and distribution of biodiversity within catchments.