Livestock grazing to maintain habitat of a critically endangered grassland bird: Is grazer species important?

Livestock grazing is an important management tool for biodiversity conservation in many native grasslands across the globe. Understanding how different grazing species interact with their environment is integral to achieving conservation goals. In the semiarid grasslands of Australia, grazing by sheep or cattle is used to manipulate vegetation structure to suit the habitat needs of a globally unique, critically endangered grassland bird, the plains-wanderer Pedionomus torquatus. However, there has been no investigation of whether sheep and cattle differ in their effects on plains-wanderer habitat and, therefore, it is unknown if these grazers are substitutable as a management tool. Using a grazing experiment in native grasslands over 3 years, we determined the effects of grazer type (sheep, cattle) on occurrence and vocal activity of plains-wanderer, vegetation structure and composition, and food availability. We also examined grazer effects on encounter rates of other grassland birds. Plains-wanderer breeding activity was inferred from vocalization rates captured by bioacoustic recorders. Spotlighting was used to measure encounter rates of other grassland birds. We found that different grazers altered the structure of the habitat. Grasslands grazed by cattle were typically more open, less variable, and lacked patches of dense vegetation relative to those grazed by sheep. Grazer type did not influence the likelihood of plains-wanderer occurrence, but it did interact with year of survey to affect breeding activity. The number of days with one or more calls significantly increased at sheep grazed sites in year-3, which coincided with enduring drought conditions. Similarly, grazer effects on encounter rate of all birds, bird species richness, and Australasian pipit Anthus novaeseelandiae were different between years. Dense vegetation specialists (such as stubble quail Coturnix pectoralis) were positively associated with grasslands grazed by sheep. As a habitat management tool, sheep or cattle grazing are useful when the goal is to support an open grassland structure for the plains-wanderer. However, their substitutability is likely to be dependent upon climate. We caution that a loss of dense vegetation in grasslands grazed by cattle during drought could limit the availability of optimal habitat for the plains-wanderer and habitat for other grassland birds.

Potassium channel modulation by a toxin domain in matrix metalloprotease 23.

Peptide toxins found in a wide array of venoms block K(+) channels, causing profound physiological and pathological effects. Here we describe the first functional K(+) channel-blocking toxin domain in a mammalian protein. MMP23 (matrix metalloprotease 23) contains a domain (MMP23(TxD)) that is evolutionarily related to peptide toxins from sea anemones. MMP23(TxD) shows close structural similarity to the sea anemone toxins BgK and ShK. Moreover, this domain blocks K(+) channels in the nanomolar to low micromolar range (Kv1.6 > Kv1.3 > Kv1.1 = Kv3.2 > Kv1.4, in decreasing order of potency) while sparing other K(+) channels (Kv1.2, Kv1.5, Kv1.7, and KCa3.1). Full-length MMP23 suppresses K(+) channels by co-localizing with and trapping MMP23(TxD)-sensitive channels in the ER. Our results provide clues to the structure and function of the vast family of proteins that contain domains related to sea anemone toxins. Evolutionary pressure to maintain a channel-modulatory function may contribute to the conservation of this domain throughout the plant and animal kingdoms.

The D-diastereomer of ShK toxin selectively blocks voltage-gated K+ channels and inhibits T lymphocyte proliferation.

The polypeptide toxin ShK is a potent blocker of Kv1.3 potassium channels, which are crucial in the activation of human effector memory T cells (T(EM)); selective blockers constitute valuable therapeutic leads for the treatment of autoimmune diseases mediated by T(EM) cells, such as multiple sclerosis, rheumatoid arthritis, and type-1 diabetes. The critical motif on the toxin for potassium channel blockade consists of neighboring lysine and tyrosine residues. Because this motif is sufficient for activity, an ShK analogue was designed based on D-amino acids. D-allo-ShK has a structure essentially identical with that of ShK and is resistant to proteolysis. It blocked Kv1.3 with K(d) 36 nm (2,800-fold lower affinity than ShK), was 2-fold selective for Kv1.3 over Kv1.1, and was inactive against other K(+) channels tested. D-allo-ShK inhibited human T(EM) cell proliferation at 100-fold higher concentration than ShK. Its circulating half-life was only slightly longer than that of ShK, implying that renal clearance is the major determinant of its plasma levels. D-allo-ShK did not bind to the closed state of the channel, unlike ShK. Models of D-allo-ShK bound to Kv1.3 show that it can block the pore as effectively as ShK but makes different interactions with the vestibule, some of which are less favorable than for native ShK. The finding that an all-D analogue of a polypeptide toxin retains biological activity and selectivity is highly unusual. Being resistant to proteolysis and nonantigenic, this analogue should be useful in K(+) channel studies; all-d analogues with improved Kv1.3 potency and specificity may have therapeutic advantages.

Targeting effector memory T cells with a selective peptide inhibitor of Kv1.3 channels for therapy of autoimmune diseases.

The voltage-gated Kv1.3 K(+) channel is a novel target for immunomodulation of autoreactive effector memory T (T(EM)) cells that play a major role in the pathogenesis of autoimmune diseases. We describe the characterization of the novel peptide ShK(L5) that contains l-phosphotyrosine linked via a nine-atom hydrophilic linker to the N terminus of the ShK peptide from the sea anemone Stichodactyla helianthus. ShK(L5) is a highly specific Kv1.3 blocker that exhibits 100-fold selectivity for Kv1.3 (K(d) = 69 pM) over Kv1.1 and greater than 250-fold selectivity over all other channels tested. ShK(L5) suppresses the proliferation of human and rat T(EM) cells and inhibits interleukin-2 production at picomolar concentrations. Naive and central memory human T cells are initially 60-fold less sensitive than T(EM) cells to ShK(L5) and then become resistant to the peptide during activation by up-regulating the calcium-activated K(Ca)3.1 channel. ShK(L5) does not exhibit in vitro cytotoxicity on mammalian cell lines and is negative in the Ames test. It is stable in plasma and when administered once daily by subcutaneous injection (10 mug/kg) attains "steady state" blood levels of approximately 300 pM. This regimen does not cause cardiac toxicity assessed by continuous EKG monitoring and does not alter clinical chemistry and hematological parameters after 2-week therapy. ShK(L5) prevents and treats experimental autoimmune encephalomyelitis and suppresses delayed type hypersensitivity in rats. ShK(L5) might prove useful for therapy of autoimmune disorders.